Tyramide Signal Amplification (TSA)

Tyramide Signal Amplification (TSA), also known as catalyzed reported deposition (CARD), offers an effective way to efficiently boost your signal and detection capabilities for low-abundance targets in IHC, ICC and FISH. HRP catalyzes the conversion of labeled inactive tyramide into a reactive radical, which then binds to adjacent tyrosine residues greatly enhancing the density of labeling. Tyramide reagents can be conjugated with a dye for fluorescent detection, or a hapten for antibody-based probing.

TSA can be used to enhance the signal of multiple targets on one sample by deactivating HRP using a peroxidase quenching buffer, then carrying out another round of TSA using a different set of antibodies from a different host species. Alternatively, the antibodies for the initial round of TSA can be stripped, leaving the covalently bound tyramides. TSA can then be repeated for the desired number of targets.

Figure 1: Tyramide Signal Amplification Principles: A primary and secondary antibody are used to label a tissue or cell sample. The secondary antibody is pre-conjugated to horseradish peroxidase (HRP), which in the presence of H₂O₂, catalyzes a labeled tyramide substrate into a highly reactive species that covalently bind to tyrosine residues on the proteins in close proximity to the antibodies and HRP, thus providing signal amplification.

Key Features of Tyramide Signal Amplification:

• Allows detection of low-abundance targets
• Used to enhance signals in IHC, ICC and FISH
• Reduces the amount of primary antibody required
• 100-fold more sensitive than conventional methods
• Simple, flexible, and easy to incorporate into IHC, IHC and FISH workflows
• Compatible with fluorescent multiplex systems

Tyramide Signal Amplification Protocol Outline for IHC and ICC

1. Follow IHC/ICC protocol for fixing, permeabilizing and blocking cells/tissue sample
2. Incubate with primary antibody
3. Incubate with HRP-conjugated secondary antibody
4. Incubate with dye-tyramide solution (e.g. Cyanine 3 tyramide (Cat. No. 6457))
5. Visualize and/or record fluorescence intensity or repeat TSA steps as necessary to label all targets (after either quenching HRP or removing antibodies)

Tyramide Signal Amplification Product Review

Find out how our customers have used and what they think of our 5 Star TSA product Biotinyl Tyramide, as they share experimental details and images of their results.

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Literature for Tyramide Signal Amplification (TSA)

Tocris offers the following scientific literature for Tyramide Signal Amplification (TSA) to showcase our products. We invite you to request* your copy today!

*Please note that Tocris will only send literature to established scientific business / institute addresses.

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